Rotational in vitro compliance measurement of diverse anastomotic configurations: a tool for anastomotic engineering

Anastomotic configurations with a small internal diameter are prone to intimal hyperplasia which can cause occlusion within weeks or months. A link between intimal hyperplasia and inhomogenities of the elastic profile of the anastomosis has been established, making anastomotic engineering directed towards smoothing the compliance profile at the anastomotic site essential. Methods to date restrict the anastomotic compliance measurement to one plane. We present a method by which the anastomotic configurations are rotated, thereby allowing an anastomotic elastic profile assessment in multiple planes. Eight end-to-end anastomoses (ovine common carotid artery) and three end-to-side anastomoses (e-PTFE graft to ovine common carotid artery) were prepared and mounted in an artificial circulation system. Anastomotic circumferential compliance (maximal-minimal diameter/(maximal-minimal pressure.minimal diameter)) was measured by means of a laser-scan-micrometer and a Statham pressure transducer. By rotating end-to-end anastomoses, the compliance was measured in three, and in end-to-side anastomoses in four different planes. Multiplanar compliance variability in areas remote to both end-to-end and end-to-side anastomoses was approximately 9%. At the suture line the variability was approximately 22% in end-to-end anastomoses and 78% in end-to-side anastomoses. These results show that local factors result in different compliance profiles when utilizing a multiplanar technique, particularly in end-to-side anastomoses. The rotational apparatus is a tool which can be used to more accurately engineer a homogeneously compliant anastomosis, with the ultimate goal of prolonging anastomotic patency.     

Experimental studies in animals on the use of a fibrin glue from the human plasma fraction Cohn I in nerve reconstruction

In order to compare the effects of suture and glue direct nerve connections and nerve transplantations in the sciatic nerve in rats were performed. 4, 6 and 12 weeks later, the nerve anastomosis were histologically studied. In the direct nerve connections, despite a single holding suture, dehiscences were frequently detected with the penetration of the adhesive between the nerve ends. Due to the exact adaptation with the aid of inserted nerve grafts, the anastomoses could be repaired sutureless successfully using a fibrin glue cuff. Since histologically, in comparison to suture, no foreign body granulomas were found, the findings in the literature could be confirmed. No cicatricial contractions of the anastomoses could be found. In the interfascicular nerve transplantation, comparatively good results may be obtained using the two-component adhesive on the Cohn I-fraction basis.     

Further technical considerations of the sleeve microanastomosis

In sleeve anastomoses, stenoses at the suture site have been the main concern. Mechanical dilatation is one way to prevent the stenosis, as suggested by Lauritzen. In the present study, 50 vessels (femoral and carotid) and 10 veins were used for sleeve anastomoses and the same numbers of vessels were used for conventional anastomoses (as control) to evaluate the effect of mechanical dilatation using resin corrosion cast (Mercox) because the Mercox cast facilitates three-dimensional stereoscopic views. Gradual dilatations around the suture sites were observed in seven carotid arteries, and three of seven resulted into aneurysm formation due to weakening of the inner vascular wall in the sleeve anastomosis. No dilatation or aneurysm was observed in the femoral arteries. Newly proliferating capillaries formed on the endothelial surfaces of the inner vascular walls around the suture sites after 4 weeks in the sleeve anastomoses. Operative time and endothelial trauma were markedly reduced with sleeve anastomoses. The gradual dilatation and aneurysm formation in the carotid arteries show that sleeve anastomoses should be used carefully for high-pressure arteries in clinical practice if mechanical dilatation is performed.     

Evaluation of anastomotic techniques in the experimental transfer of free skin flaps.

Vascularized groin flaps were transferred to the contralateral side by various microvascular anastomotic techniques. There was no statistical difference between the results of end-to-end and end-to-side anastomoses, when the same sized vessels were compared. However, significant differences existed when vessels smaller than 0.5 mm in diameter were anastomosed.     

Preferential use of end-to-side arterial anastomoses in free flap transfers.

In our early clinical experience with free flaps, we used end-to-end arterial anastomoses and in 9 our of 24 we had complete failures--7 of which were due to early arterial thrombosis. Contrarily, in 41 consecutive free flaps with end-to-side anastomoses we have not had a single failure. At the same time that we began using the end-to-side anastomoses, we also began using the latissimus dorsi free flap as our flap of first choice, and we agree that this was probably an additional reason for our improved success rate. The use of end-to-side anastomoses has the following advantages: (1) a high success rate; (2) preservation of all existing vessels in an injured extremity; (3) greater freedom of operative planning; and (4) technical simplicity in terms of access to the vessels. For us, these advantages have made end-to-side anastomosis the technique of choice in the transfer of free flaps.     

Experimental study of results of nerve suture under tension vs. nerve grafting.

Evoked electromyograms and myelinated fiber caliber histograms were used to compare the results of end-to-end nerve suture and nerve grafting, performed with an operating microscope. The best results were achieved after direct suture without tension. Direct suture under a tension of 25 grams produced results sometimes comparable the that of the zero tension group, but mostly quite inferior to those and even to the grafted group. The regeneration of axons in the nerve graft group was delayed more than in the sutured groups, but by the tenth month the number of regenerating fibers had increased with stable and comparatively good results. After a nerve suture under a tension of 25 grams, there is impairment of the blood flow at the suture site, and the results are not as good. In such situations I believe nerve grafting should be performed. This amount of tension also seems to be the limit which will allow accurate coaptation of a nerve by microsurgical techniques.     

Microvascular anastomoses in irradiated vessels: a comparison between the Unilink system and sutures

A new mechanical device (the Unilink system) was compared to conventional suture anastomoses in irradiated microvessels. Twenty rabbits received a single radiation dose of 20 Gy from a 7-MeV electron source through an anterior neck field. One and 6 months following irradiation, the carotid arteries and facial veins were divided and anastomosed on one side with the Unilink system and on the other side with suture technique. At sacrifice 4 weeks postoperatively, all vessels were evaluated for patency and histologic changes associated with radiation and anastomotic trauma. Histology disclosed severe radiation changes. Also, intimal hyperplasia was consistently found at the anastomotic sites in the arteries, while it was totally absent in the venous anastomoses. Occlusive thrombosis was found in two arteries, one anastomosed with the Unilink system and one sutured. Two other arteries, one from each group, had subtotal occlusions at the anastomotic site. No occlusions occurred in any of the venous anastomoses. The overall patency in this study was 97.5 percent, with no difference between the two techniques.     

Testing of a new prototype surgical stapler that automates the rollover sleeve technique for venous anastomoses

The creation of successful vascular anastomoses is of primary importance in many surgical fields. Numerous attempts to automate this process have been made. These techniques have slowly gained acceptance, but their use is still limited. This report details feasibility testing of a new prototype stapler that automates the rollover sleeve technique for venous vascular anastomoses. Male and female mongrel dogs (n=7) (25-32 kg) were used. A segment of the right (n=5) or left (n=2) iliac vein was harvested for interposition grafts after the contra lateral side was transected. In each dog, two end-to-end venous anastomoses at the interposition grafts were performed. The standard anastomosis employed continuous mattress sutures. The experimental anastomosis was performed with a new prototype surgical stapler. The stapled anastomosis was proximal and the sutured was distal. In all experiments, it was possible to perform the experimental anastomosis with the stapler. Complications included two small leaks, one due to misfiring of a single pin in one experimental site. These leaks required suture reinforcement. One dog died of hemorrhage due to a slipped suture at the vein harvest site. One vein had thrombus seen at the sutured site although no technical abnormalities at either of the anastomoses could be found. After two weeks, grafts were inspected grossly and histologically. Healing appeared normal. There was a trend for less inflammatory cells infiltrating stapled sites; however, this was not statistically significant. The experiments demonstrate that this device can automate the rollover sleeve technique for venous anastomoses.     

Vascular anastomosis using controlled phase transitions in poloxamer gels

Vascular anastomosis is the cornerstone of vascular, cardiovascular and transplant surgery. Most anastomoses are performed with sutures, which are technically challenging and can lead to failure from intimal hyperplasia and foreign body reaction. Numerous alternatives to sutures have been proposed, but none has proven superior, particularly in small or atherosclerotic vessels. We have developed a new method of sutureless and atraumatic vascular anastomosis that uses US Food and Drug Administration (FDA)-approved thermoreversible tri-block polymers to temporarily maintain an open lumen for precise approximation with commercially available glues. We performed end-to-end anastomoses five times more rapidly than we performed hand-sewn controls, and vessels that were too small (<1.0 mm) to sew were successfully reconstructed with this sutureless approach. Imaging of reconstructed rat aorta confirmed equivalent patency, flow and burst strength, and histological analysis demonstrated decreased inflammation and fibrosis at up to 2 years after the procedure. This new technology has potential for improving efficiency and outcomes in the surgical treatment of cardiovascular disease.     

Video microsurgery: evaluation of standard laparoscopic equipment for the practice of microsurgery

Traditional microsurgery involves the use of bulky and expensive stereo microscopes that have limited portability. Recent advances in video technology have enabled the exploration of alternative visualization methods. The purpose of this study was to evaluate standard laparoscopic equipment for microvascular anastomoses. Eight surgeons completed anastomoses on rat femoral and synthetic vessels using stereo microsurgery and video microsurgery visualization systems. All surgeons had previous experience with stereo microsurgery and none had ever used video microsurgery. Data were collected on overall anastomosis and individual suture times. A sample of completed anastomoses was placed in a video database and evaluated by use of a quality rating scale (8 to 10, excellent; 6 to 7, adequate; less than 6, poor). All surgeons subjectively evaluated the video microsurgery system. A total of 48 anastomoses were completed. The average total anastomosis time for the stereo microsurgery was 1018.9 +/- 463.2 seconds versus 1738.9 +/- 460.1 seconds for the video microsurgery. The average individual suture placement time was 114.6 +/- 60.6 seconds for the stereo microsurgery versus 211.7 +/- 128.4 seconds for the video microsurgery (p < 0.05). Twenty-five of the anastomoses underwent quality review. The overall score of the stereo microsurgery group was 8.1 +/- 1.7, and the video microsurgery group had an overall score of 7.3 +/- 1.6. Survey results revealed that 75 percent of the participants thought that the video microsurgery would be useful for human operations and would improve surgeon comfort, but 87.5 percent would not use the present video microsurgery system over stereo microsurgery in their practice. Although significant differences exist in overall anastomosis and individual suture completion times, no difference was found in the overall quality. Video microsurgery could become a useful tool on the basis of surgeon ergonomics; however, optical parameters require further refinement.     

End-to-end versus end-to-side microvascular anastomosis patency in experimental venous repairs

In this experimental study, venous end-to-end and end-to-side microvascular anastomoses in similar and diameter-discrepant vessels were compared. In 50 rats, end-to-end microvascular repair of the divided epigastric vein and end-to-side repair of the epigastric vein into the femoral vein showed 5-day patency rates of 75 and 88 percent, respectively. These data are not statistically different. In 20 rats, microvascular repair of end epigastric to end femoral veins (size discrepant) and end epigastric to side femoral veins showed 5-day patency rates of 50 and 85 percent, respectively. These data are statistically different (p less than 0.05). We conclude from these experimental data that end-to-side venous repairs may be useful in lowering the anastomosis thrombosis rate seen when size-discrepant veins are repaired.     

Sciatic neuropathy following endovascular treatment of a limb vascular malformation

Background and aim

Allow for protection of briefly ischemic tissues against the harmful effects of subsequent prolonged ischemia is a phenomennon called as Ischemic Preconditioning (IP). IP has not been studied in ischemia-reperfusion (I/R) model of peripheral nerve before. We aimed to study the effects of acute IP on I/R injury of peripheral nerve in rats.

Method

70 adult male rats were randomly divided into 5 groups in part 1 experimentation and 3 groups in part 2 experimentation. A rat model of severe nerve ischemia which was produced by tying iliac arteries and all idenfiable anastomotic vessels with a silk suture (6-0) was used to study the effects of I/R and IP on nerve biochemistry. The suture technique used was a slip-knot technique for rapid release at time of reperfusion in the study. Cytoplasmic vacuolar degeneration was also histopathologically evaluated by light microscopic examination in sciatic nerves of rats at 7th day in part 2 study.

Results

3 hours of Reperfusion resulted in an increase in nerve malondialdehyde levels when compared with ischemia and non-ischemia groups (p < 0.001 and p < 0.0001 respectively). IP had significantly lower nerve MDA levels than 3 h reperfusion group (p < 0.001). The differences between ischemic, IP and non-ischemic control groups were not significant (p > 0.05). There was also a significant decrease in vacoular degeneration of sciatic nerves in IP group than I/R group (p < 0.05).

Conclusion

IP reduces the severity of I/R injury in peripheral nerve as shown by reduced tissue MDA levels at 3 th hour of reperfusion and axonal vacoulization at 7 th postischemic day.     

THE TIME COURSE OF SIEVE TUBE AND VESSEL REGENERATION AND THEIR RELATION TO PHLOEM ANASTOMOSES IN MATURE INTERNODES OF COLEUS

Quantitative counts of regenerative sieve tubes and vessels were made in a large number of samples of mature internode #5 of C. blumei, with concomitant study of the fine details of vascular regeneration and the occurrence of the normally developing phloem anastomoses. Such anastomoses were found in many of the plants, but their average number in the small regenerating area was low (viz., 0.9 ± 0.2). With the phloem anastomoses excluded from the counts, the time course of regeneration was clear cut—no strands completed their regeneration around the wound until three days after wounding. More regenerative sieve tubes completed their differentiation under all conditions than did regenerative vessels. The number of sieve tubes and vessels regenerated by four days was closely related to the number of preexisting bundles of that type of vascular cell that had been severed by the transverse wound. The ratio of bundles severed by the wound in the phloem to those in the xylem was 2.14, and the ratio of the regenerative sieve tubes to the regenerative vessels was 2.24. For both tracheary and sieve tube cells the initial regeneration was strongly polar (mostly above the wound), as expected from earlier IAA transport data. The path of tracheary regeneration was obviously related to that of the sieve tubes on the other side of the cambium.     

Reconstruction of the Abdominal Vagus Nerve Using Sural Nerve Grafts in Canine Models

Background

Recently, vagus nerve preservation or reconstruction of vagus has received increasing attention. The present study aimed to investigate the feasibility of reconstructing the severed vagal trunk using an autologous sural nerve graft.

Methods

Ten adult Beagle dogs were randomly assigned to two groups of five, the nerve grafting group (TG) and the vagal resection group (VG). The gastric secretion and emptying functions in both groups were assessed using Hollander insulin and acetaminophen tests before surgery and three months after surgery. All dogs underwent laparotomy under general anesthesia. In TG group, latency and conduction velocity of the action potential in a vagal trunk were measured, and then nerves of 4 cm long were cut from the abdominal anterior and posterior vagal trunks. Two segments of autologous sural nerve were collected for performing end-to-end anastomoses with the cut ends of vagal trunk (8–0 nylon suture, 3 sutures for each anastomosis). Dogs in VG group only underwent partial resections of the anterior and posterior vagal trunks. Laparotomy was performed in dogs of TG group, and latency and conduction velocity of the action potential in their vagal trunks were measured. The grafted nerve segment was removed, and stained with anti-neurofilament protein and toluidine blue.

Results

Latency of the action potential in the vagal trunk was longer after surgery than before surgery in TG group, while the conduction velocity was lower after surgery. The gastric secretion and emptying functions were weaker after surgery in dogs of both groups, but in TG group they were significantly better than in VG group. Anti-neurofilament protein staining and toluidine blue staining showed there were nerve fibers crossing the anastomosis of the vagus and sural nerves in dogs of TG group.

Conclusion

Reconstruction of the vagus nerve using the sural nerve is technically feasible.     

Innervation of the arteriovenous anastomoses in the dog tongue

Summary Profiles of nerve plexuses in the arteriovenous anastomoses of the dog tongue were investigated by both transmission and scanning electron microscopy. Three-dimensional morphology of the vascular nerves was examined after removal of the connective tissue components by the HCl-hydrolysis method. Tight bending and a rich nerve supply were the most characteristic features of the anastomosing channels. The tunica media consisted of an outer circular layer of typical smooth-muscle cells and an inner region containing longitudinal plicae of ramified smoothmuscle cells. The tunica adventitia was exclusively occupied by nerve bundles; fibroblasts were poorly developed. Numerous nerve bundles of variable size were coiled around the anastomosing channels, and occasional bundles ran crosswise over the U-shaped bent vessels.     

A histologic comparison of experimental microarterial end-in-end (sleeve) and end-to-end anastomoses

Twenty-five microvascular end-in-end and 21 end-to-end anastomoses were performed on the central arteries (diameters 0.8 to 1.2 mm) of rabbit ears. The appearance of longitudinal specimens taken after intervals of 1 to 2 hours and 1, 3, 7, 14, 30, and 90 days was studied microscopically. Between 1 to 2 hours and 14 days, all end-in-end anastamoses showed considerable narrowing of the telescoped segment, and platelets had accumulated at the sleeved vessel end. The vascular walls of the telescoped segment were thickened distally, and the internal elastic laminas were folded in the lingitudinal direction due to contraction. Extensive loss of endothelium was observed both around sutures and distally in the telescoped segments. The stenosis initially noted in telescoped vessels gradually decreased due to atrophy of the medial layers in the double vascular walls. Following the organization of the platelet aggregates, subsequent endothelialization was completed between 7 and 14 days. After 2 weeks, neointimal hyperplasia was frequently found at the sleeved vessel end. In the early postoperative period, platelets covered exposed subendothelial structures and sealed defects in the vessel walls of all end-to-end anastomoses. During the first few postoperative weeks, the medial layers disintegrated and were replaced by increasing fibrosis. Reendothelialization was completed between 7 and 14 days, whereafter gradual intimal hyperplasia became significant.(ABSTRACT TRUNCATED AT 250 WORDS)     

Experimental reclamping of free-flap pedicles: the effect of prolonged stasis on the anastomoses and clamp sites

The effect of static blood in direct contact with areas of microvascular anastomoses and previous clamp application for prolonged periods of time has been investigated. The free groin flap was used as a model in 27 white rabbits. The flap pedicle vessels were reclamped proximal to the anastomoses and areas of previous clamp application for periods of time varying between 30 minutes and 4 hours after 15 minutes of blood flow over these areas. A 100 percent patency rate was achieved despite the long periods of reclamping. Histologically, minor intimal damage was visible in the immediate period following anastomoses and clamping of the vessels. After 2 weeks, despite a thickened myofibroblastic intimal lesion, an intact endothelial layer was observed. No evidence of thrombosis could be demonstrated in either period. We postulate that vessels carefully treated and with technically well-performed anastomoses can be regarded as "normal" vessels after 15 minutes of blood flow over the anastomoses and clamp sites. We suggest that when required, microvascular clamps may then be reapplied without risk for prolonged periods of time despite static blood being in contact with these areas.     

Vascular Anastomosis—Sutures,Staples or Glue?

Different vascular suturing techniques were assessed in the experimental laboratory. Staples made of tantulum wire were employed for vascular anastomoses in normal dogs'' arteries and veins and in atherosclerotic rabbits'' aortas. The staples were driven with the NRC Vascular Suturing Instrument. In a similar series of experiments a plastic bonding agent (Eastman 910) was used. The anastomosed vessels were found to be patent and few complications were encountered. However, it is unlikely that the conventional suture technique with the currently available materials will be replaced by these methods except under special circumstances. Clinical application of the stapling method is envisaged in the anastomosis of small vessels and in situations when speed is essential, whereas the bonding agent promises to be useful as an ancillary method in combination with sutures or staples.     

The use of prostacyclin analogue-containing suture for the prevention of postoperative venous thrombosis in the rat

The ability of prostacyclin analogue incorporated into a controlled-release suture to prevent postoperative venous thrombosis was investigated. Thirteen rats underwent bilateral transection and anastomosis of the common femoral vein. In each animal, polycaprolactone suture containing 0.25 micrograms/cm of the prostacyclin analogue Iloprost (Schering Ag, Berlin, West Germany) was used to perform the anastomosis on one vessel. Similar suture without prostacyclin analogue was used on the contralateral vessel, which served as a control. Functional patency and luminal surface morphology were assessed 24 hours postoperatively. All anastomoses performed using suture containing prostacyclin analogue were patent. Among controls, five anastomoses were patent and eight were occluded. This difference was highly significant (p less than 0.005). All anastomoses performed with prostacyclin analogue-containing suture exhibited a uniform absence of thrombosis. In contrast, eight control veins exhibited a dense, well-organized fibrinous clot that filled the entire lumen, effectively sealing off the vessel. These results suggest that the prostacyclin analogue released from the suture was highly effective in inhibiting thrombus formation without adversely affecting the vessel's ability to achieve hemostasis.     

Continuous sutures for microarterial end-to-side anastomoses

A continuous suture technique for end-to-side microarterial anastomoses has been developed and tested in an experimental model on rat common carotid arteries.